Nucleic acid analysis is used in a variety of clinical and research applications. Traditionally, a separate nucleic acid (NA) analysis step was performed by hand in a bench top setting, which requires relatively large amounts of material, labor, and time. For example, in order to undergo nucleic acid analysis, a sample must first be prepped for analysis. Whole blood samples must first be processed to separate red blood cells from the sample. In other samples, for instance, a respiratory sample there may be fewer or no processing steps. Nonetheless, the sample must be lysed, purified, amplified, and then detected, all in separate, manual steps. These steps are labor-intensive and also traditionally require varied and expensive laboratory equipment. There is a need to scale down nucleic acid analysis and detection into an automated device that is capable of performing all the various steps in NA analysis.
Previous CD platforms have accomplished lysing by combining cell samples with grinding media (milling beads) and exposing the system to intense mixing by rapid and abrupt motion of the container. Lysis initially occurred due to collisions between milling beads and cells, causing puncture by direct collision and friction based shearing. However, in this system a secondary lysis mechanism took place when alternating the spin directions, causing impaction and friction due to Coriolis effects in the CD chamber. Another CD system improved on this by replacing the Coriolis-induced lysis with stronger magnetic-assisted bead beating. For example, U.S. Patent Application Publication No. 2007-0125942 discloses examples of this. Generally, strong magnets placed loosely in the CD were actuated by permanent magnets in a stationary platform causing additional impaction and lysis in the radial direction. The main limitation of the stand-alone systems remained the possibility for integration with other processing steps.
This invention combines elements of both Coriolis induced lysis and magnetically assisted lysis to create a superior CD-based lysis system that is integrated with additional processing steps.